ROCK通路在取向纳米纱纤维环组织工程支架内BMSC细胞形变与定向分化中的作用与机制

项目名称： ROCK通路在取向纳米纱纤维环组织工程支架内BMSC细胞形变与定向分化中的作用与机制

项目编号： No.81472071

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 叶晓健

作者单位： 中国人民解放军第二军医大学

项目金额： 80万元

中文摘要： 椎间盘突出术后复发与纤维环破损及无法有效修复密切相关。组织工程技术可望有效修复，但对材料适配度以及材料界面细胞生长分化的诱导、调控及其机制的掌握将决定修复的成效。申请者前期制备的取向纳米纱支架(AYS)，具有类纤维环胞外基质结构和良好力学强度，在与骨髓间充质干细胞(BMSC)的三维培养中发现BMSC变形并长入支架内。因ROCK信号通路与细胞骨架蛋白相关，可调节细胞形变并参与TGF-β和CTGF对细胞功能的调控，本项目拟通过干预ROCK通路探索AYS内BMSC形变的机制；用TGF-β和CTGF诱导BMSC分化的同时通过调节ROCK通路，观察该通路在BMSC定向分化中的作用。在猪椎间盘纤维环缺损模型中，移植入不同调控状态下的AYS-BMSC复合体，观察界面融合效果，验证ROCK通路在AYS和生长因子共同诱导BMSC向纤维环细胞分化中的作用，为组织工程修复纤维环提供必要的理论依据和实验基础。

中文关键词： 纤维环；组织工程支架；ROCK信号通路；骨髓间充质干细胞；定向分化

英文摘要： The high postoperative recurrence of intervertebral disc herniation usually results from defects of annulus fibrosus (AF) and limited repairing potential. It is proposed that tissue-engineering technology can effectively repair the ruptured AF. However, the repairing effect depends on the material compatibility, inducing and regulation of interfacial cell growth differentiation, and the mastery of its mechanism. We have fabricated a novel aligned nanoyarn scaffold（AYS） which mimics the structure of AF extracellular matrix and has good mechanical strength. The bone marrow mesenchymal stem cells (BMSCs) are found to be deformed and grow into the scaffold in three-dimensional cell culture. Because ROCK signal pathway is associated with cytoskeletal protein, it can regulate cell deformation and participate in the regulation of TGF-β and CTGF on cell functions. This study aims to investigate the mechanism of morphologic changes of BMSC inside AYS through the intervention of ROCK pathway and observe the effect of ROCK pathway regulation in the directed differentiation of BMSC induced by TGF-β and CTGF. We implanted different AYS-BMSC composites into the model of defected pig AF and observed its interface fusion effect, so as to verify the function and mechanism of ROCK pathway in BMSC differentiation induced jointly by AYS and exogenous growth factors. The study will provide necessary theoretical and experimental basis for AF repair using tissue engineering technology.

英文关键词： annulus fibrosus;tissue engineering scaffold;ROCK signaling pathway;bone marrow mesenchymal stem cell;committed differentiation